Method for Obtaining Information on Formation of Double-Stranded Nucleic Acid

Abstract

Provided is a method for obtaining information on the formation of a double-stranded nucleic acid by detecting fluorescence, which is simple, is applicable to various interaction systems, and features high detection sensitivity with reduced background signals. Specifically provided is a method for obtaining information on the formation of a double-stranded nucleic acid, which includes detecting information on fluorescence of a probe consisted of a labeling fluorescent dye labeled on a nucleic acid strand and an intercalator bound or inserted between base pairs in the double-stranded nucleic acid to permit an energy transfer with the fluorescent dye.

1 . A method for obtaining information on formation of a double-stranded nucleic acid between a first nucleic acid strand and a second nucleic acid strand having a base sequence complementary to that of the first nucleic acid strand, which comprises detecting information on fluorescence of a probe consisted of a labeling fluorescent dye labeled on the first nucleic acid strand and an intercalator bound or inserted between base pairs in the double-stranded nucleic acid to permit an energy transfer with the fluorescent dye. 
     
     
         2 . The method according to  claim 1 , wherein the probe is comprised of a group of plural fluorescent dyes including at least an excitation-target fluorescent dye capable of giving excitation energy, which has been obtained by absorption of light, to another fluorescent dye and a detection-target fluorescent dye capable of fluorescing by receiving the excitation energy from the another fluorescent dye. 
     
     
         3 . The method according to  claim 1 , wherein the intercalator is a quencher that quenches the labeling fluorescent dye. 
     
     
         4 . The method according to  claim 1 , wherein the first nucleic acid strand is immobilized at an end thereof on a surface of a region of interaction. 
     
     
         5 . A method for obtaining information on hybridization, which comprises conducting at least:
 a step of allowing the hybridization to proceed between a first nucleic acid strand, on which a fluorescent dye is labeled, and a second nucleic acid strand having a base sequence complementary to that of the first nucleic acid strand,   a step of adding an intercalator which can be bound or inserted between base pairs in the double-stranded nucleic acid to permit an energy transfer with the fluorescent dye, and   a step of detecting information on fluorescence of the fluorescent dye and intercalator.   
     
     
         6 . A method for obtaining information on an amount of a nucleic acid strand having a base sequence complementary to that of a probe nucleic acid strand, which comprising detecting information on fluorescence of a probe consisted of a labeling fluorescent dye labeled on the probe nucleic acid strand and an intercalator bound or inserted between base pairs in a double-stranded nucleic acid to permit an energy transfer with the fluorescent dye.